A | B | |
1x, ul | ||
oligo-dT, 10uM | 1 | |
dNTP, 10mM | 1 |
A | B | C | D | |
1x, ul | 4x, ul | 12.2x, ul | ||
5x SmartScribe Buffer | 2 | 8 | 24.4 | |
DTT, 100mM | 1 | 4 | 12.2 | |
TSO primer, 10uM | 0.3 | 1.2 | 3.66 | |
H2O | 1.45 | 5.8 | 17.69 | |
Superase-In | .25 | 1 | 3.05 | |
SmartScribe RT (add last) | 1 | 4 | 12.2 |
A | B | C | |
1x, ul | 12.2x, ul | ||
KAPA Hifi HotStart | 12.5 | 152.5 | |
ISPCR primers, 10uM | 1 | 12.2 | |
RNaseA | .75 | 9.15 | |
Lambda Exonuclease | .75 | 9.15 |
A | B | |
ul | ||
Pooled cDNA | x | |
Splint | 10-x | |
2x NEBuilder Assembly Mix | 10 |
A | B | C | D | |
1x, ul | 3.2x, ul | 6.2x | ||
NEB Buffer 2 | 5 | 16 | 31 | |
H2O | 16 | 51.2 | 99.2 | |
Exonuclease I | 3 | 9.6 | 18.6 | |
Exonuclease III | 3 | 9.6 | 18.6 | |
Lambda exonuclease | 3 | 9.6 | 18.6 |
A | B | C | |
1x, ul | 3.2x, ul | ||
Phi29 buffer | 5 | 16 | |
dNTP (10 mM) | 2.5 | 8 | |
Random hexamer primers (exo resistant) | 2.5 | 8 | |
H2O | 29 | 92.8 | |
Phi29 polymerase | 1 | 3.2 |